HBV DNA Test Among Blood Donations May Require Two Amplification Targets

Area of Science:

• Virology
• Blood Transfusion Safety
• Molecular Diagnostics

Background:

• Hepatitis B virus (HBV) DNA testing is crucial for blood donation safety.
• Commercial nucleic acid test (NAT) reagents may yield false-negative results, impacting transfusion safety.
• Understanding the reasons for false negatives is essential for improving HBV screening.

Purpose of the Study:

• To analyze the risks and causes of false-negative HBV DNA results from commercial NAT reagents in Chinese blood donations.
• To evaluate the necessity of using two amplification targets for HBV DNA detection in blood donations.

Main Methods:

• Discordant samples from two commercial NAT platforms were re-tested using an in-house method targeting conserved HBV genome regions.
• High-titer samples were analyzed by the in-house method, with discordant region results sequenced and aligned with primers/probes.
• HBV DNA concentrations were quantified using commercial reagents and compared to in-house findings.

Main Results:

• The in-house method demonstrated good detection performance.
• Primer-template mismatches in conserved S and C regions affected detection capacity but could be compensated by other regions.
• While most false negatives were due to low titers, 7 samples with high HBV DNA concentrations (above LOD) indicated potential primer/probe mismatches in commercial reagents.

Conclusions:

• Commercial NAT reagents carry a risk of false-negative HBV DNA detection.
• Dual-target assays are recommended for HBV screening to mitigate the risk of false-negative results in blood donations.